PKC lambda/iota CRISPR Activation Plasmid (m): sc-422263-ACT

Prkci encodes the atypical protein kinase C isoform PKC lambda/iota, a serine/threonine kinase that functions as a core component of the PAR polarity complex and regulates apical–basal polarity, asymmetric cell division, and tight junction organization. In mouse cells, PKC lambda/iota integrates upstream cues from small GTPases and phosphoinositide signaling to coordinate cytoskeletal dynamics, vesicle trafficking, and directional migration. It modulates proliferative and survival signaling through pathways that intersect with PI3K–AKT, NF-κB, and Hippo/YAP transcriptional outputs, influencing differentiation and tissue architecture. Dysregulation of atypical PKC activity has been associated with altered epithelial integrity, aberrant growth control, and inflammation-linked phenotypes, making Prkci a useful node for mechanistic studies in development, regeneration, and disease models.

PKC lambda/iota CRISPR Activation Plasmid (m) provides a targeted, non-destructive approach to upregulating endogenous Prkci expression without altering the underlying DNA sequence.

PKC lambda/iota CRISPR Activation Plasmid (m) is a three-plasmid synergistic activation mediator (SAM) system engineered for highly efficient, site-specific transcriptional upregulation of the Prkci locus in human cell lines. The system is built around a catalytically inactive Cas9 (dCas9) carrying two inactivating mutations (D10A and N863A) that eliminate nuclease activity while preserving DNA binding. This dCas9 is fused to VP64, a potent transcriptional activator, and is co-expressed with a blasticidin resistance gene for selection. The second plasmid encodes the MS2-p65-HSF1 fusion protein, a secondary activator complex that works in concert with dCas9-VP64, alongside a hygromycin resistance gene. The third plasmid encodes a target-specific 20 nt sgRNA fused to two MS2 RNA aptamers that recruit the MS2-p65-HSF1 complex to the activation site, accompanied by a puromycin resistance gene. The three plasmids are delivered at a 1:1:1 mass ratio for balanced expression of all system components.

Once assembled at the target locus, the SAM complex binds within approximately 200 bp upstream of the Prkci transcriptional start site, where VP64, p65, and HSF1 act in concert to recruit transcriptional machinery and drive upregulation of endogenous PKC lambda/iota expression. Unlike nuclease-active Cas9, dCas9 does not introduce double-strand breaks or modify the genomic sequence, preserving the native Prkci locus and enabling the study of PKC lambda/iota-dependent transcriptional responses at the endogenous locus, making it a valuable tool for functional studies, target gene identification, and the modeling of PKC lambda/iota pathway restoration in tumor cells with silenced or reduced Prkci expression.

For Research Use Only. Not Intended for Diagnostic or Therapeutic Use.